GenScript can also synthesize desalt and HPLC purified guide RNA compatible with saCas9, Cas12f/CasMINI, Cas13, and other Cas nuclease variants. Click here to enter your guide sequence (target sequence + scaffold sequence) and order online. Testimonials...
The guide RNA sequences were cloned into the dual-CRISPR-Cas9-U6-H1 (pBP48) plasmid containing the guide RNA scaffold and Cas9 sequence. K562 cells were transfected with the plasmids containing the respective pairs of guide RNAs and then selected for successful transfection using puromycin. K562 ...
The crRNA is usually a 17–20 nucleotide sequence complementary to the target DNA, and the tracrRNA serves as a binding scaffold for the Cas9 nuclease. While crRNAs and tracrRNAs exist as two separate RNA molecules in nature, the single-guide RNA (sgR...
DNA templates for sgRNAs based on an optimized scaffold were made24 and transcribed in vitro into sgRNAs using T7 RNA polymerase. The sgRNA pools were purified with VAHTS RNA Clean Beads, aliquoted, and stored at −80 °C. T7 RNA polymerase was ordered from ThermoScientific (Cat. no....
essential for separating cells from the tissue scaffold and ensuring the production of a single-cell suspension. (3) Cell population enrichment. This step employs techniques to isolate specific target cells from the heterogeneous cell suspension. Methods such as FACS or MACS can be utilized to ...
To test the activity of each Cpf1-family protein, we selected a guide RNA target site within the DNMT1 gene (Figure 7B). We first found that each of the Cpf1-family proteins along with its respective crRNA designed to target DNMT1 was able to cleave a PCR amplicon of the DNMT1 ...
essential for separating cells from the tissue scaffold and ensuring the production of a single-cell suspension. (3) Cell population enrichment. This step employs techniques to isolate specific target cells from the heterogeneous cell suspension. Methods such as FACS or MACS can be utilized to ...
Current single-cell RNA sequencing (scRNA-seq) methods with high cellular throughputs sacrifice full-transcript coverage and often sensitivity. Here we describe Smart-seq3xpress, which miniaturizes and streamlines the Smart-seq3 protocol to substantially reduce reagent use and increase cellular throughpu...
Eliminate any read that maps to an unassembled scaffold, mitochondrial DNA, or any locus that is marked as a repeat element by RepeatMasker (http://www.repeatmasker.org). The repeat elements by RepeatMasker were retrieved from UCSC genome browser [30], with the exception of opossum, where we...
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